Page 268 - Color Atlas of Biochemistry

Genetic engineering 259

A. Restriction endonucleases

H H Palindrome
H H
3' G 5' G A A T T C 3'
O H
3' C T T A A G 5'
O P O H C 5' O
2
EcoRI DNA-Ligase
O H
A 3.1.21.4 6.5.1.1
H 3'
5' G A A T T C 3'
H
C T T A A G
H H 3' 5'
3' G
OH H
O A A T T C 3'
5' G
O P O H C 5' O G 5'
2
O A H 3' C T T A A EcoRI + DNA
3'
Overhanging ends

B. DNA cloning

DNA
G AA TT C G AA TT C
C TT AA G C TT AA G

EcoRI
EcoRI
Purification Vector DNA
(Plasmid)

Isolated DNA ligase
DNA fragment

Host cell Gene for AATTC
(bacterium) antibiotic G
resistance
CTTAA G
Recombinant
plasmid

Transformation

Plasmid
Plasmid
isolation
Cleavage
Bacterial with
genome
EcoRI

Bacterial culture in the Cloned
presence of antibiotic DNA fragment

Koolman, Color Atlas of Biochemistry, 2nd edition © 2005 Thieme
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